Ecdysteroids fractionation

Fig. 4.4. HPLC fractionation of the free ecdysteroid fraction from Moniezia expansa (a) sample 1, 3.3% portion (bj sample 2, 10% portion on a chromatographic system with collection of fractions every minute for the radioimmunoassay (RIA) (ICT-1 antiserum). The positions of authentic ecdysteroids are shown IV, 20, 26-dihydroxyecdysone II, 20-hydroxyecdysone I, ecdysone. (After Mendis et al., 1984.)...

The analyses thus far have only been conducted with female pupae and are incomplete and the scheme of isolation is shown in Figure 3. The butanol phase which contained the free ecdysteroids was further fractionated by counter-current distribution (CCD) [60 transfers in a system of cyclohexane-butanol-water (4 6 10) with 10 ml of each phase]. The material from CCD tubes 23-42 was subjected to three Cie SEP-PAK fractionations (13). Quantitative RP-HPLC and radioassay analyses of fractions T and 5 resulting from using a SEP-PAK elution system as in Figure 3 yielded 200 Mg of labeled 20-hydroxyecdysone. The combined residues from CCD tubes 43-57 when fractionated over a Florisil SEP-PAK cartridge and chromatographed over 4 g of Silica gel yielded 22 pg of labeled ecdysone. 

Fig. 2.9. DCCC separation of ecdysteroids from Bombyx mori (silkworm) ethyl acetate extract (0.65 g), using CHCI3-MeOH-H2O (13 7 4, v/v), ascending mode, 1.0ml per fraction, detection at 254 nm (221)...

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